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Effects of the Surface Densities of Glycoclusters on the Determination of Their IC 50 and K d Value Determination by Using a Microarray
Author(s) -
Dupin Lucie,
Zuttion Francesca,
Géhin Thomas,
Meyer Albert,
PhanerGoutorbe Magali,
Vasseur JeanJacques,
Souteyrand Eliane,
Morvan François,
Chevolot Yann
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500371
Subject(s) - chemistry , biofilm , lectin , microarray , pseudomonas aeruginosa , high throughput screening , nanotechnology , bacteria , biochemistry , biology , materials science , genetics , gene expression , gene
Pseudomonas aeruginosa (PA) is an opportunistic bacterium involved in 10–30 % of nosocomial diseases. It causes severe lung injury to cystic fibrosis patients, often leading to patient death. PA strains are multidrug resistant, thus making the design of new therapeutics a challenge for public health. One promising therapeutic option is to design glycoclusters that target the virulence factor of PA. LecA is a galactose‐specific lectin that might be involved in adhesion and biofilm formation by PA. The DNA‐directed immobilization (DDI) microarray is a powerful tool for screening and understanding of structure–activity relationships between glycoclusters and lectins. High‐throughput and multiplexed analysis of lectin–glycocluster interactions on a DDI microarray allows measurement of IC 50 and dissociation constant ( K d ) values with minute amounts of material. In order to study the robustness of the DDI microarray in determination of IC 50 and K d values, the impact of glycocluster surface density was investigated. The data obtained show that measured IC 50 values were influenced by glycocluster surface density: as the density of glycoclusters increases, the measured IC 50 values increase too. In contrast, the measured K d values were not affected by glycocluster surface density, provided that the experimental conditions allow interaction between glycocluster and lectin at single‐molecule level (no surface cluster effect).